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The avascular nature of hyaline cartilage results in limited spontaneous self-repair and regenerative capabilities when damaged. Recent advances in three-dimensional bioprinting have enabled the precise dispensing of cell-laden biomaterials, commonly referred to as 'bioinks', which are emerging as promising solutions for tissue regeneration. An effective bioink for cartilage tissue engineering needs to create a micro-environment that promotes cell differentiation and supports neocartilage tissue formation. In this study, we introduced an innovative bioink composed of photocurable acrylated type I collagen (COLMA), thiol-modified hyaluronic acid (THA), and poly(ethylene glycol) diacrylate (PEGDA) for 3D bioprinting cartilage grafts using human nasal chondrocytes. Both collagen and hyaluronic acid, being key components of the extracellular matrix (ECM) in the human body, provide essential biological cues for tissue regeneration. We evaluated three formulations - COLMA, COLMA+THA, and COLMA+THA+PEGDA - for their printability, cell viability, structural integrity, and capabilities in forming cartilage-like ECM. The addition of THA and PEGDA significantly enhanced these properties, showcasing the potential of this bioink in advancing applications in cartilage repair and reconstructive surgery.
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Ácido Hialurónico , Ingeniería de Tejidos , Andamios del Tejido , Ácido Hialurónico/química , Ácido Hialurónico/farmacología , Ingeniería de Tejidos/métodos , Humanos , Andamios del Tejido/química , Condrocitos/citología , Condrocitos/efectos de los fármacos , Polietilenglicoles/química , Bioimpresión/métodos , Colágeno/química , Impresión Tridimensional , Cartílago/citología , Matriz Extracelular/química , Supervivencia Celular/efectos de los fármacos , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , TintaRESUMEN
Processing cellulose from 4-methyl morpholine n-oxide (NMMO)-water solutions is a completely circular route that produces biodegradable cellulose fibers or films while recovering reusable NMMO [Guo, Y.; Cai, J.; Sun, T.; Xing, L.; Cheng, C.; Chi, K.; Xu, J.; Li, T. The purification process and side reactions in the N-methylmorpholine-N-oxide (NMMO) recovery system. Cellulose 2021, 28(12), 7609-7617]. Despite proven success in two-dimensional applications, challenges in transitioning to three-dimensional objects arise from the critical changes that cellulose undergoes during deposition, regeneration, and postregeneration stages. While emphasizing the critical diffusion-driven precipitation during regeneration, this investigation explores the influence of extrusion temperature, printing alignment, regeneration, and drying processes on interfilament fusion, bonding, shape integrity, and mechanical properties. Three distinct drying processes: ambient, vacuum, and freeze-drying were investigated. Tensile and flexural bending tests provided insight into the delamination of dried specimens. Ambient and vacuum drying enhanced the properties of specimens, while freeze-drying resulted in a more stable shape. The findings contribute to advancing the understanding of 3D-printing cellulose from NMMO solutions, addressing crucial aspects of the extrusion, regeneration, and drying stages for enhanced applications in sustainable manufacturing.
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Celulosa , Impresión Tridimensional , Agua , Celulosa/química , Agua/química , Óxidos N-Cíclicos/química , Morfolinas/química , Soluciones , Desecación/métodosRESUMEN
Geopolymers are alkaline-activated aluminosilicate binders recognized as a promising alternative to traditional Portland cement due to their significantly lower greenhouse emissions, energy consumption, and carbon footprint. However, the challenge is meeting or exceeding the strength of Portland cement concrete while being prepared within a desired setting time and possessing workable rheology. A "water-in-air" Pickering emulsion, also called dry water, was prepared by stabilizing water droplets with hydrophobic nano silica and using them to control the geopolymer's strength, setting time, and workability. The mechanisms that underlie the effects of dry water on the rheology, setting, and strength were studied in detail through a combination of rheological, thermal, morphological, chemical, and microstructural assessments. A reduction in the viscosity and yield shear stress manifests in a higher flow diameter, principally due to the particle size coarsening in the precursor and the flowability of hydrophobic nano silica. There was a rapid rise in temperature during the setting process as the dry water temporarily increased the local alkalinity in the mixture, which boosted the dissolution of the precursor and, hence, the reaction. Outcomes from X-ray diffraction, thermogravimetric analysis, and Fourier-transform infrared confirm the highest degree of polycondensation for the principal N-A-S-H framework in mixtures containing dry water. These eventually correspond to a denser microstructure under scanning electron microscopy and, in turn, a superior mechanical strength. Depending on the unique combination of characteristics, including size coarsening, temporary water encapsulation, microfilling effect, and supplementary silica source, dry water resolves the "trade-off" between geopolymer's fresh and hardened properties when introducing nanoparticles.
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Hydrogels appear to be an attractive class of biomaterial for cartilage tissue engineering due to their high water content, excellent biocompatibility, tunable stiffness, etc. The crosslinking density of the hydrogel can affect their viscoelastic property, and therefore potentially impact the chondrogenic phenotype of re-differentiated chondrocytes in a 3D microenvironment through physical cues. To understand the effect of crosslinking densities on chondrocytes phenotype and cellular interaction with the hydrogel, this study utilized a clinical grade thiolate hyaluronic acid and thiolate gelatin (HA-Gel) hydrogel, crosslinked with poly(ethylene glycol) diacrylate to create various crosslinking densities. The HA-Gel hydrogels were then mixed with human nasal chondrocytes to generate neocartilage in vitro. The influence of the hydrogel crosslinking density and the viscoelastic property on the cell behaviours on the gene and matrix levels were evaluated using biochemistry assays, histology, quantitative polymerase chain reaction (qPCR) and next-generation sequencing (RNA seq). In general, the differences in the storage modulus of the HA-Gel hydrogel are not enough to alter the cartilaginous gene expression of chondrocytes. However, a positively correlated trend of PPAR-γ gene expression to the crosslinking density was measured by qPCR. The RNA-seq results have shown that 178 genes are significantly negatively correlated and 225 genes are positively correlated to the crosslinking density, which is worth investigating in the future studies.
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Due to the limited self-repair capacity of the hyaline cartilage, the repair of cartilage remains an unsolved clinical problem. Tissue engineering strategy with 3D bioprinting technique has emerged a new insight by providing patient's personalized cartilage grafts using autologous cells for hyaline cartilage repair and regeneration. In this review, we first summarized the intrinsic property of hyaline cartilage in both maxillofacial and orthopedic regions to establish the requirement for 3D bioprinting cartilage tissue. We then reviewed the literature and provided opinion pieces on the selection of bioprinters, bioink materials, and cell sources. This review aims to identify the current challenges for hyaline cartilage bioprinting and the directions for future clinical development in bioprinted hyaline cartilage.
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This article aims to understand the rheology of collagen networks and their role in various stages of a bioprinting process while building tissue-like constructs. The science of rheology, which deals with the deformation and flow of matter, has grown considerably from its earlier focus on polymer melts and solutions and their processing methods to hydrogels with new processing procedures, such as bioprinting. The main objective of this paper is to discuss the impact of the rheology of collagen hydrogels on micro-extrusion and layer-stacking stages of bioprinting. Generally, the rheological characterization of hydrogels, including collagens by dynamic measurements under small deformations, is considered sufficient to evaluate their bioprinting performance. However, we brought out the importance of other rheological properties of collagen networks, such as steady-state shear flow conditions and large amplitude oscillator shear. While the dynamic measurements under small deformations help characterize the crosslinking and gel formations of the collagen, the steady shear flow measurements are better tools for investigating filament micro-extrusion and layer-stacking stages of a bioprinting process. We brought the role of other non-Newtonian material functions, such as first normal stress difference and extensional viscosity in addition to shear viscosity, for the first time. Extensional viscosity and the viscoelasticity manifested through normal-stress differences are significant in capillary (needle) flow. We also suggested caution to use dynamic viscosity vs. oscillation frequency under small deformations in place of steady shear viscosity vs. shear rate measurement. In addition, we brought out the importance of the large amplitude oscillatory shear test to investigate the collagen networks under large deformations. Finally, we discussed the role of crosslinking and flow conditions on cell viability. Those discussions are focused on collagen networks; nevertheless, they are valid on the bioprinting of other hydrogels.
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Bioimpresión , Bioimpresión/métodos , Reología , Hidrogeles , Viscosidad , ColágenoRESUMEN
The interaction between compositional ratios, namely, SiO2/Al2O3, Na2O/Al2O3, H2O/Na2O and the liquid-to-solid ratio, triggers mutual sacrifice between workability, setting time and strength for N-A-S-H geopolymers. The present study characterizes the mechanism underlying the effect of these compositional ratios and, in turn, develops guidelines for mixture design that requires a simultaneous and satisfactory delivery of these engineering properties. The experimental results show that an increase in either the SiO2/Al2O3, Na2O/Al2O3 or H2O/Na2O ratio raises the liquid-to-solid ratio, which in turn improves the workability of fresh mixtures. A continuous increase beyond 2.8 for the SiO2/Al2O3 ratio boosts its strength, but also significantly extends its final set. Lowering the Na2O/Al2O3 ratio from 1.3 to 0.75 raises the compressive strength significantly, while the shortest final set was seen at the median value, 1.0. A H2O/Na2O ratio of 9~10 yields the highest strength and the fastest final set simultaneously, due to the maximized degree of geopolymerization. Moreover, the accompanying sensitivity analysis indicates that the workability depends chiefly upon the H2O/Na2O ratio, the final setting time on the SiO2/Al2O3 ratio and, that the compressive strength relies on both of them. Also, this study proposes an optimal range of 2.8~3.6 for SiO2/Al2O3, 0.75~1.0 for Na2O/Al2O3 and 9~10 for H2O/Na2O to guarantee high strength, together with high flow and within the allowable final setting time. Furthermore, multi-factor predictive models are established with acceptable accuracy for practitioners to regulate oxide compositions in N-A-S-H geopolymers, which will guide future mixture design.
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Over the past few years, nanocellulose and its derivatives have drawn attention as promising bio-based materials for water treatment applications due to their high surface area, high strength, and renewable, biocompatible nature. The abundance of hydroxyl functional groups on the surfaces of cellulose nanocrystals (CNCs) and cellulose nanofibrils (CNFs) enables a broad range of surface modifications which results in propitious nanocomposites with tunable characteristics. In this context, this review describes the continuously developing applications of nanocellulose-based materials in the areas of adsorption, catalysis, filtration, and flocculation, with a special emphasis on the removal of contaminants such as heavy metals, dyes, and pharmaceutical compounds from diverse water systems. Recent progresses in the diverse forms of application of nanocellulose adsorbents (suspension, hydrogel, aerogel, and membrane) are also highlighted. Finally, challenges and future perspectives on emerging nanocellulose-based materials and their possible industrial applications are presented and discussed.
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Nanocompuestos , Nanopartículas , Purificación del Agua , Celulosa/química , Hidrogeles/química , Nanopartículas/química , Purificación del Agua/métodosRESUMEN
The removal of skin cancer lesions on the nose often results in the loss of nasal cartilage. The cartilage loss is either surgically replaced with autologous cartilage or synthetic grafts. However, these replacement options come with donor-site morbidity and resorption issues. 3-dimensional (3D) bioprinting technology offers the opportunity to engineer anatomical-shaped autologous nasal cartilage grafts. The 3D bioprinted cartilage grafts need to embody a mechanically competent extracellular matrix (ECM) to allow for surgical suturing and resistance to contraction during scar tissue formation. We investigated the effect of culture period on ECM formation and mechanical properties of 3D bioprinted constructs of human nasal chondrocytes (hNC)-laden type I collagen hydrogel in vitro and in vivo. Tissue-engineered nasal cartilage constructs developed from hNC culture in clinically approved collagen type I and type III semi-permeable membrane scaffold served as control. The resulting 3D bioprinted engineered nasal cartilage constructs were comparable or better than the controls both in vitro and in vivo. This study demonstrates that 3D bioprinted constructs of engineered nasal cartilage are feasible options in nasal cartilage reconstructive surgeries.
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We investigated the gelation and microstructure of cellulose nanocrystals (CNCs) in nonionic hydroxyethyl cellulose (HEC) solutions. Cellulose nanocrystals (CNCs) with a particle length of 90 nm and width of 8 nm currently produced by acid hydrolysis of wood pulp were used in this study. The microstructures of CNCs/polymer suspensions were investigated by performing linear small amplitude oscillatory shear (SAOS) and nonlinear large amplitude oscillatory shear (LAOS), in addition to constructing CNCs phase diagrams and measuring steady-state shear viscosities. Significant viscosity increases at low shear rates coupled with high shear thinning behaviors were observed in CNCs in HEC solutions above the overlapping concentration of HEC. The physical strength of CNCs/HEC solution gels increased with the increase in CNCs concentration and resembled the weakly crosslinked gels according to the scaling of linear dynamic mechanical experiments. According to LAOS analysis, CNCs/HEC mixtures showed type III behavior with intercycle stress softening, while the samples showed stress stiffening in single cycles.
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Objective: The avascular inner regions of the knee menisci cannot self-heal. As a prospective treatment, functional replacements can be generated by cell-based 3D bioprinting with an appropriate cell source and biomaterial. To that end, human meniscus fibrochondrocytes (hMFC) from surgical castoffs of partial meniscectomies as well as cellulose nanofiber-alginate based hydrogels have emerged as a promising cell source and biomaterial combination. The objectives of the study were to first find the optimal formulations of TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl)-oxidized cellulose nanofiber/alginate (TCNF/ALG) precursors for bioprinting, and then to use them to investigate redifferentiation and synthesis of functional inner meniscus-like extracellular matrix (ECM) components by expanded hMFCs. Methods: The rheological properties including shear viscosity, thixotropic behavior recovery, and loss tangent of selected TCNF/ALG precursors were measured to find the optimum formulations for 3D bioprinting. hMFCs were mixed with TCNF/ALG precursors with suitable formulations and 3D bioprinted into cylindrical disc constructs and crosslinked with CaCl2 after printing. The bioprinted constructs then underwent 6 weeks of in vitro chondrogenesis in hypoxia prior to analysis with biomechanical, biochemical, molecular, and histological assays. hMFCs mixed with a collagen I gel were used as a control. Results: The TCNF/ALG and collagen-based constructs had similar compression moduli. The expression of COL2A1 was significantly higher in TCNF/ALG. The TCNF/ALG constructs showed more of an inner meniscus-like phenotype while the collagen I-based construct was consistent with a more outer meniscus-like phenotype. The expression of COL10A1 and MMP13 were lower in the TCNF/ALG constructs. In addition, the immunofluorescence of human type I and II collagens were evident in the TCNF/ALG, while the bovine type I collagen constructs lacked type II collagen deposition but did contain newly synthesized human type I collagen.
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The fabrication of fiber mats via electrospinning has been adopted in the last decades to produce high quality scaffolds for tissue engineering. However, an effective combination of electrospinning methods with gene delivery therapies remains a challenge. In this study, we describe how the delivery of gene complexes via electrospun mats that contain different volumes of gelatin (Gel), collagen (Col), and polyethylene glycol (PEG) can affect gene expression by transfected cells. Non-viral complexes were formulated by using lipid modified polyethylenimine (PEI) polymer and plasmid DNAs (pDNA) like the reporter Green Fluorescent Protein (GFP) and the therapeutically relevant Bone Morphogenetic Protein-2 (BMP-2) and electrospuned after being mixed with different volumes of Gel-Col-PEG mats and delivered to human myoblast (C2C12) and mouse osteoblast cells (MC3T3). The entrapment of GFP complexes via different homogeneous electrospun fiber mats revealed that a high fraction of collagen in the mats affected the quality of the fibers and led to reduced transfection efficiency on target cells. On the other hand, the fabrication of double-layered mats that contained collagen without complexes as a first layer and gelatin-collagen-PEG with complexes as a second layer successfully induced GFP expression and ALP activity in C2C12 cells. We conclude that this study has established the advantage of formulating multilayered bioactive collagen-based mats for gene delivery applications.
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We investigated the gelation of cellulose nanocrystals (CNCs) in polyelectrolyte and neutral polymer solutions. Cellulose nanocrystals (CNCs) with half-ester sulfate groups produced by acid hydrolysis of wood pulp were used in this study. The microstructure of CNCs/polymer suspensions was investigated in semidilute concentration regimes by selecting carboxymethyl cellulose (CMC700) as an anionic polymer and poly(ethylene oxide) (PEO600) as a neutral polymer solution. Together with quartz crystal microbalance with dissipation monitoring (QCM-D), rheology, scanning electron microscopy (SEM), and cryo-transmission electron microscopy (cryo-TEM), we characterized CNCs-polymer interactions, the suspension microstructure, and the macroscopic gel flow. Significant viscosity increases at low shear rates coupled with high shear-thinning behaviors were observed in CNC colloid-CMC700 polymer mixtures, but not those CNCs in PEO600 solutions. The apparent differences between CNCs-CMC700 and CNCs-PEO600 mixtures were due to their chain confirmations. On the basis of the evaluations from STEM, cryo-TEM, and polarized optical microscopy, we proposed that the excess CMC700 molecules in solutions result in the depletion of CNCs and the formation of anisotropic domains.
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Skin cancer patients often have tumorigenic lesions on their noses. Surgical resection of the lesions often results in nasal cartilage removal. Cartilage grafts taken from other anatomical sites are used for the surgical reconstruction of the nasal cartilage, but donor-site morbidity is a common problem. Autologous tissue-engineered nasal cartilage grafts can mitigate the problem, but commercially available scaffolds define the shape and sizes of the engineered grafts during tissue fabrication. Moreover, the engineered grafts suffer from the inhomogeneous distribution of the functional matrix of cartilage. Advances in 3D bioprinting technology offer the opportunity to engineer cartilages with customizable dimensions and anatomically shaped configurations without the inhomogeneous distribution of cartilage matrix. Here, we report the fidelity of Freeform Reversible Embedding of Suspended Hydrogel (FRESH) bioprinting as a strategy to generate customizable and homogenously distributed functional cartilage matrix engineered nasal cartilage. Using FRESH and in vitro chondrogenesis, we have fabricated tissue-engineered nasal cartilage from combining bovine type I collagen hydrogel and human nasoseptal chondrocytes. The engineered nasal cartilage constructs displayed molecular, biochemical and histological characteristics akin to native human nasal cartilage.
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Bioimpresión/métodos , Cartílago Articular/citología , Condrocitos/química , Colágeno/química , Hidrogeles/química , Tabique Nasal/citología , Ingeniería de Tejidos/métodos , Adulto , Cartílago Articular/fisiología , Condrogénesis , Humanos , Masculino , Andamios del Tejido/químicaRESUMEN
In the current study, we focused on the preparation of nanocomposite films of natural rubber latex-cellulose nanocrystals (NR/CNCs) and investigated the release of CNCs from those materials into aqueous solutions. The obtained nanocomposite films were well characterized for further understanding of the release mechanism; as the intermolecular interactions between the two components were studied by Fourier-transform infrared spectroscopy, the morphology was studied with scanning electron microscopy, and nanostructures were analyzed by tensile and dynamic mechanical testing. The release behavior of CNCs from the NR/CNCs nanocomposite films was studied by a fluorescent labeling technique, and the release process in various media was modeled by first-order kinetics. Higher contents of CNCs in the nanocomposite films and a relatively acidic or alkaline medium facilitated the release process, while higher ionic strength of the media could hamper the release of CNCs from the nanocomposite films. In this study, our objective was to transport CNC particles from NR/CNC composites into immersed media to be used beneficially in biomedical applications. Nevertheless, in other surroundings, the release of CNCs or any other nanoparticles from composite materials may not be desirable. Hence, this study also provides a protocol to investigate the release of nanoparticles from a host matrix into the surrounding media and also promotes a rethinking of the nanoparticle release issue from composites to the environment.
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Celulosa/química , Látex/química , Nanopartículas/química , Ensayo de Materiales , Microscopía Electrónica de Transmisión , Estructura Molecular , AguaRESUMEN
A huge variety of plants are harvested worldwide and their different constituents can be converted into a broad range of bionanomaterials. In parallel, much research effort in materials science and engineering is focused on the formation of nanoparticles and nanostructured materials originating from agricultural residues. Cellulose (40-50%), hemicellulose (20-40%), and lignin (20-30%) represent major plant ingredients and many techniques have been described that separate the main plant components for the synthesis of nanocelluloses, nano-hemicelluloses, and nanolignins with divergent and controllable properties. The minor components, such as essential oils, could also be used to produce non-toxic metal and metal oxide nanoparticles with high bioavailability, biocompatibility, and/or bioactivity. This review describes the chemical structure, the physical and chemical properties of plant cell constituents, different techniques for the synthesis of nanocelluloses, nanohemicelluloses, and nanolignins from various lignocellulose sources and agricultural residues, and the extraction of volatile oils from plants as well as their use in metal and metal oxide nanoparticle production and emulsion preparation. Furthermore, details about the formation of activated carbon nanomaterials by thermal treatment of lignocellulose materials, a few examples of mineral extraction from agriculture waste for nanoparticle fabrication, and the emerging applications of plant-based nanomaterials in different fields, such as biotechnology and medicine, environment protection, environmental remediation, or energy production and storage, are also included. This review also briefly discusses the recent developments and challenges of obtaining nanomaterials from plant residues, and the issues surrounding toxicity and regulation.
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Nanoestructuras , Aceites Volátiles , Celulosa , Lignina , PolisacáridosRESUMEN
A semidilute cellulose nanocrystal suspension was tested for pressure, volume, temperature dependencies of its viscosity and density. The compression of a 2.0 wt % cellulose nanocrystal suspension under 5.0 MPa at room temperature resulted in morphological changes from istotropic to nematic form. However, at high temperature, high-pressure treatment caused desulfation and gelation. Those results have significant applications, not only as additives in drilling and fracturing fluids but also for the preparation of hydrogels.
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Celulosa/química , Calor , Hidrogeles/química , Nanopartículas/química , Presión , Agua/química , Fuerza Compresiva/fisiología , Calor/efectos adversos , Presión/efectos adversos , Suspensiones , ViscosidadRESUMEN
Calcium hydroxide is used in diverse applications including heritage conservation where supplying it in the form of nanoparticles allows easy carbonation with atmospheric air contacts. The effects of cellulose nanofibers on the precipitation of calcium hydroxide nanoparticles were investigated by varying the reaction time, concentration, and carboxylation content of cellulose nanofibers. Cellulose nanofibers were very effective in producing calcium hydroxide nanoparticles with less than 50 nm sizes out of calcium nitrate-sodium hydroxide precipitation reactions. The formation of smaller-size calcium hydroxide nanoparticles is believed to be the result of heterogeneous nucleation and growth of calcium hydroxide particles on cellulose nanofibers. The liquid-phase nucleated and grown calcium hydroxide nanoparticles were also deposited onto cellulose nanofibers. The resulting calcium hydroxide nanoparticles were carbonized and generated calcite under atmospheric carbon dioxide in an efficient way.
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TEMPO-oxidized cellulose nanofibers (TOCNFs) were prepared and investigated as low-cost adsorbents for the removal of Cu (II) and Zn (II) from synthetic and natural waters. The adsorption equilibrium was reached within 2â¯min and adsorption capacity was as high as 102.9â¯mgâ¯g-1 for Cu (II) and 73.9â¯mgâ¯g-1 for Zn (II). Ionic strength showed adverse effect on adsorption capacity, however, TOCNFs with higher carboxymethyl content were less influenced due to their resistance to aggregation. Copper adsorption exhibited strong selectivity over the tested common cations. The adsorptions of Cu (II) and Zn (II) onto TOCNF were endothermic but spontaneous processes, and the binding was driven by entropy increase. A combined interaction mechanism, including ion exchange, coordination and accumulation, was proposed based on the study. All the findings confirmed the great potential of TOCNF application in water purification and reclamation approaches.
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Extrusion-based additive manufacturing (EBAM) or 3D printing is used to produce customized prototyped parts. The majority of the polymers used with EBAM show moisture sensitivity. However, moisture effects become more pronounced in polymers used for critical applications, such as biomedical stents, sensors, and actuators. The effects of moisture on the manufacturing process and the long-term performance of Shape Memory Polyurethane (SMPU) have not been fully investigated in the literature. This study focuses primarily on block-copolymer SMPUs that have two different hard/soft (h/s) segment ratios. It investigates the effect of moisture on the various properties via studying: (i) the effect of moisture trapping within these polymers and the consequences when manufacturing; (ii) and the effect on end product performance of plasticization by moisture. Results indicate that higher h/s SMPU shows higher microphase separation, which leads to an increase of moisture trapping within the polymer. Understanding moisture trapping is critical for EBAM parts due to an increase in void content and a decrease in printing quality. The results also indicate a stronger plasticizing effect on polymers with lower h/s ratio but with a more forgiving printing behavior compared to the higher h/s ratio.